Taxonomy of an anamorphic xylariaceous fungus from a termite nest found together with <Emphasis Type="Italic">Xylaria angulosa</Emphasis>

Two xylariaceous fungi were isolated from a nest of a termite, Odontotermes formosanus, that was incubated in a laboratory after collecting from Iriomote Is., Okinawa Pref., in Japan. One of the two fungi was identified as Xylaria angulosa on the basis of the morphology of branched stroma produced on medium, tiny asci, and ascospores having a germ slit. Another fungus is an anamorphic fungus that produces synnemata up to 50 mm long from which dendritic conidiophores branch out. Unicellular conidia are holoblastically produced on a sympodially proliferating conidiogenous cell. Such morphological characters resemble those of the genus Geniculosporium. However, its distinctive synnema formation and dendritic conidiophores do not assign the fungus to Geniculosporium or other known genera and warrant establishment of a new genus. The phylogenetic tree based on the ITS regions of rDNA shows that the fungus is nested in the cluster of the genus Nemania (Xylariaceae), whose species have mainly Geniculosporium-like anamorphs. We describe here the present anamorphic fungus as Geniculisynnema termiticola gen. et sp. nov., and discuss its phylogenetic and ecological relationships to xylariaceous fungi, especially termiticolous species.     

Phylogenetic affiliation of the desert truffles <Emphasis Type="Italic">Picoa juniperi</Emphasis> and <Emphasis Type="Italic">Picoa lefebvrei</Emphasis>

The molecular phylogeny and comparative morphological studies reported here provide evidence for the recognition of the genus Picoa, an hypogeous desert truffle, in the family Pyronemataceae (Ascomycota, Pezizales). Picoa juniperi and Picoa lefebvrei were reassigned to the genus Picoa based on large subunit (LSU) sequence (28S) rDNA and internal transcribed spacer (ITS) rDNA (including the partial 18S, ITS1, ITS2, 5.8S gene, and partial 28S of the nuclear rDNA) data. Morphological studies of spores, asci, perida, and gleba revealed high similarities between P. lefebvrei and P. juniperi, thereby confirming the membership of both species in the genus Picoa. These two species were primarily distinguishable based on ascospore ornamentation.     

Conidial development in selected ambrosial species of the genus <Emphasis Type="Italic">Raffaelea</Emphasis>

Conidiogenesis of the type species and two additional species of the genus Raffaelea Arx & Hennebert are described. In contrast to previous conclusions based on light microscopy that indicate sympodial production of the conidia, we found that conidium development was by annellidic percurrent proliferation in the type species Raffaelea ambrosiae, as well as in Raffaelea arxii and Raffaelea albimanens. Consequently, this mode of conidium development is similar to the conidiogenesis of anamorphs within the Ophiostomatales, but distinguished by sporodochia formed within the genus Raffaelea. These findings reduce the differences between Raffaelea and other anamorphs with annellidic percurrent proliferation of the conidiogenous cells within the Ophiostomatales to different conidiomata and conidiophores, and they support previous reports of the close phylogenetic relationship between Raffaelea and Ophiostoma.     

Phylogenetic placement of <Emphasis Type="Italic">Marasmiellus juniperinus</Emphasis>

Recent collections and the type specimen of Marasmiellus juniperinus, the type species of the genus, were examined. Phylogenetic placement, based on ribosomal large subunit (LSU) and internally transcribed spacer (ITS) sequences, is within the lentinuloid clade, nested among Gymnopus taxa. This placement dictates genus name usage and phylogenetic position of other putative species of Marasmiellus. The mating system is tetrapolar.     

Teleomorph-anamorph relationships and reclassification of <Emphasis Type="Italic">Cordyceps cuboidea</Emphasis> and its allied species

Based on morphological characteristics and molecular phylogeny, we reclassified Cordyceps cuboidea and allied species C. alboperitheciata, C. prolifica, and Ophiocordyceps ryogamiensis. We investigated their teleomorph-anamorph relationships and revealed that these four species have Hirsutella-like anamorphs with morphological differences between them. By analyzing their molecular phylogeny, inferred from DNA sequences of internal transcribed spacer (ITS) and large subunit (LSU) D1/D2 region of rDNA, they were separated into four close-knit clades. Although C. prolifica and O. ryogamiensis formed their own clades, isolates of C. cuboidea separated into two clades, i.e., a true C. cuboidea clade and one resembling a new species, the O. paracuboidea clade. The latter two species are distinguished by the fruiting region of the stroma. In addition, C. alboperitheciata is regarded as a synonym of C. cuboidea. From the morphology, teleomorph-anamorph relationships, and molecular phylogeny, we concluded these species should be assigned to the genus Ophiocordyceps.     

ITS rDNA sequence comparisons resolve phylogenetic relationships in <Emphasis Type="Italic">Orostachys</Emphasis> subsection <Emphasis Type="Italic">Appendiculatae</Emphasis> (Crassulaceae)

Orostachys (Crassulaceae) is a small genus of succulent plants having a predominantly East Asian distribution. Recent DNA sequence comparisons revealed polyphyletic nature of the genus and found distant relationship between its infrageneric taxa. Here we present the first molecular phylogeny of Orostachys subsection Appendiculatae based on a large number of ITS rDNA sequences representing most currently recognized members of the subsection and utilizing secondary structure information. Ribosomal spacer was a highly informative marker and provided a phylogenetic signal sufficient to resolve relationships at different scales, from affinities between species to a fine geographic structure in broadly sampled species. It was also conservative enough to allow unambiguous alignment and construction of consensus secondary structure models for ITS1 and ITS2. These models displayed a number of molecular synapomorphies defining most lineages established in our analyses. We revealed a major split in the subsection placing three species, O. spinosa, O. japonica and O. chanetii, into a strongly supported clade to the exclusion of O. thyrsiflora. Phenotypically distinct monotypic genus Meterostachys was also resolved as a part of the subsection’s clade and showed affinity to O. thyrsiflora. Our data suggested that morphology-based species concept for O. thyrsiflora requires reassessment.     

<Emphasis Type="Italic">Sativumoides</Emphasis> and <Emphasis Type="Italic">Cladosporiopsis</Emphasis>, two new genera of hyphomycetes from China

Sativumoides punicae gen. et sp. nov. and Cladosporiopsis ovata gen. et sp. nov. are described and illustrated. Sativumoides is characterized by macronematous, individual, percurrently extending conidiophores, integrated, monoblastic, apical, lageniform conidiogenous cells, and single, apical, pale brown to brown, garlic-bulb-shaped dictyosporous conidia with predominantly longitudinal septa. Cladosporiopsis is unique in possessing pigmented, apiosporous conidia seceding schizolytically from integrated, monoblastic, determinate conidiogenous cells; the conidia, arising from the sides or ends of ramoconidia, form short, acropetal chains. Sativumoides is compared with similar anamorphic genera including Triposporium, Iyengarina, Acrodictys, Actinocladium, Piricaudiopsis, Cheiroidea, Stephanoma, Domingoella, Acrogenospora and Shrungabeeja. The distinctions between Cladosporiopsis and similar genera including Cladosporium, Ochrocladosporium, Rachicladosporium, Rhizacladosporium, Toxicocladosporium, Ramularia, Bispora, Lylea and Devriesia are also discussed. Keys to these two novel genera and their morphologically similar genera are provided.     

Two new species of <Emphasis Type="Italic">Trichoderma</Emphasis> from Yunnan,China

Two new species of the fungal genus Trichoderma, Trichoderma compactum and Trichoderma yunnanense, isolated from rhizosphere of tobacco in Yunnan Province, China are described based on morphological characters and phylogenetic analyses of nucleotide sequences. Our DNA sequences included the internal transcribed spacer (ITS) regions of the rDNA cluster (ITS1 and ITS2), and partial sequences of the translation elongation factor 1-alpha (tef1) and a fragment of the gene coding for endochitinase 42 (ech42). The analyses show that T. compactum belongs to the Harzianum clade, and T. yunnanense belongs to the Hamatum clade.     

Phylogenetic relationships in genus <Emphasis Type="Italic">Arachis</Emphasis> based on ITS and 5.8S rDNA sequences

Background  

The genus Arachis comprises 80 species and it is subdivided into nine taxonomic sections (Arachis, Caulorrhizae, Erectoides, Extranervosae, Heteranthae, Procumbentes, Rhizomatosae, Trierectoides, and Triseminatae). This genus is naturally confined to South America and most of its species are native to Brazil. In order to provide a better understanding of the evolution of the genus, we reconstructed the phylogeny of 45 species using the variation observed on nucleotide sequences in internal transcribed spacer regions (ITS1 and ITS2) and 5.8 S of nuclear ribosomal DNA.     

Reclassification of Two <Emphasis Type="Italic">Peronospora</Emphasis> Species Parasitic on <Emphasis Type="Italic">Draba</Emphasis> in <Emphasis Type="Italic">Hyaloperonospora</Emphasis> Based on Morphological and Molecular Phylogenetic Data

On the family Brassicaceae, the causal agent responsible for downy mildew disease was originally regarded as a single species, Peronospora parasitica (now under Hyaloperonospora), but it was recently reconsidered to consist of many distinct species. In this study, 11 specimens of Peronospora drabae and P. norvegica parasitic on the genus Draba were investigated morphologically and molecularly. Pronounced differences in conidial sizes (P. drabae: 14–20 × 12.5–15.5 μm; P. norvegica: 20–29 × 15.5–22 μm) and 7.8% sequence distance between their ITS1-5.8S-ITS2 rDNA sequences confirmed their status as distinct species. Based on ITS phylogeny and morphology (monopodially branching conidiophores, flexuous to sigmoid ultimate branchlets, hyaline conidia and lobate haustoria), the two species unequivocally belong to the genus Hyaloperonospora and not to Peronospora to which they were previously assigned. Therefore, two new combinations, Hyaloperonospora drabae and H. norvegica, are proposed. The two taxa are illustrated and compared using the type specimen for H. norvegica and authentic specimens for H. drabae, which is lectotypified.     

Phylogeny of Chinese <Emphasis Type="Italic">Polystichum</Emphasis> (Dryopteridaceae) based on chloroplast DNA sequence data (<Emphasis Type="Italic">trnL-F</Emphasis> and <Emphasis Type="Italic">rps4-trnS</Emphasis>)

Polystichum is one of the largest and most taxonomically complex fern genera in China. The evolutionary relationships of Chinese Polystichum and related genera, and the relationship between our Polystichum phylogeny and ecogeographic distribution, were tested by the use of DNA sequence data. Fifty-one species of Polystichum and 21 species in allied genera were sequenced for the plastid intergenic spacers rps4-trnS and trnL-F. Maximum parsimony and Bayesian phylogenetic analyses of both individual and combined data sets showed that Chinese Polystichum as commonly recognized was paraphyletic: one clade (the CCPC clade) included Cyrtomidictyum lepidocaulon, two Cyrtogonellum species, three Cyrtomium species, and a small number of Polystichum species usually occurring on limestone. A second clade, Polystichum sensu stricto, included the remainder of the Polystichum species; these often occur on non-limestone substrates. The remaining Cyrtomium species formed the third clade. Three subclades resolved within Polystichum sensu stricto (s.s.) clade do not correspond with recent sectional classifications, and we outline the issues relevant to a new classification for the genus. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Molecular Systematic Study of <Emphasis Type="Italic">Chrysosplenium</Emphasis> Series <Emphasis Type="Italic">Pilosa</Emphasis> (Saxifragaceae) in Korea

To address systematic issues pertaining to series Pilosa of Korean Chrysosplenium, the phylogenetic relationships of the 35 accessions of Korean Chrysosplenium taxa were examined using the DNA sequence data obtained from the nuclear ribosomal internal transcribed spacer (ITS) regions and the psbK-I gene of the plastid genome. The results from both ITS and psbK-I sequence data indicated that Chrysosplenium flaviflorum formed a well-supported clade that merits species status of the taxon. The ITS phylogenetic trees detected two distinct lineages in Chrysosplenium pilosum var. fulvum. Geographically, one lineage was widely distributed throughout the country whereas the other was confined to Gyeonggi and Gangwon provinces. The plants sampled from Jeju Island, which have been recognized as C. pilosum var. sphaerospermum [C. hallaisanense] exhibited no DNA sequence feature segregating them from C. pilosum var. fulvum [C. barbatum] distributed in the mainland area of Korea. The plastid DNA sequence data was not fully in concordance with the nuclear ribosomal ITS data and the current taxa delimitation. These results suggest incomplete lineage sorting or that plastid genome capture may have occurred during the evolution of some taxa examined in the present study.     

<Emphasis Type="Italic">Capitulocladosporium clinodiplosidis</Emphasis> gen. et sp. nov., a hyphomyceteous ustilaginomycete from midge

A hyphomyceteous fungus producing light-colored ramoconidia was isolated from midge (Clinodiplosis sp.) in southern China. In this study, this fungus was compared with 16 morphologically similar Cladosporium-like genera, but could not be assigned to any of them due to its distinct morphological characteristics, namely absence of sympodial conidiogenous cells and presence of swollen head-like conidiogenous regions. Moreover, on the basis of phylogenetic analyses of 18S ribosomal DNA (rDNA), 5.8S rDNA, and 28S rDNA, as well as genes for the RNA polymerase II subunit and the translation elongation factor 1-alpha, this fungus was identified as a member of the Ustilaginomycetes in Basidiomycota. Therefore, Capitulocladosporium clinodiplosidis gen. et sp. nov. is proposed to accommodate the new taxon, and its morphology is described and illustrated herein. The new genus is placed in Ustilaginomycetes but without further specification on order and family level (incertae sedis), since its teleomorph and lifestyle are currently unclear.     

Phylogenetic relationships of the anamorphic genus <Emphasis Type="Italic">Fusicladium</Emphasis> s. lat. as inferred by ITS nrDNA data

The genus Fusicladium s. lat. (incl. Pollaccia and Spilocaea) was phylogenetically analysed using ITS nrDNA sequences. Pollaccia and Spilocaea did not form monophyletic groups of their own, but were intermingled between Fusicladium species, together with which they formed a monophyletic clade. Thus, Pollaccia and Spilocaea should be included in a wider genus concept of Fusicladium, constituting a morphologically variable genus. Furthermore, all Venturia and Fusicladium isolates clustered together on the bases of available ITS data, providing support for the monophyly of the anamorphic genus Fusicladium and the teleomorphic genus Venturia. Within this clade several subclades can be recognized. All taxa on the host family Salicaceae were found in one subclade. Three other subclades comprised taxa on Rosaceae whereas taxa on other host families all clustered separately. Geographic specializations were not observed. Two examples of host switching could be demonstrated, but these were confined to instances involving host species belonging to the same family. Fusicladium convolvularum and F. effusum, two species with unknown teleomorphs, clustered within the Fusicladium/Venturia clade, supporting the correct placement of these taxa in Fusicladium. The placement of Pseudocladosporium hachijoense within the family Venturiaceae was also supported.     

<Emphasis Type="Italic">Umbelopsis gibberispora</Emphasis> sp. nov. from Japanese leaf litter and a clarification of <Emphasis Type="Italic">Micromucor ramannianus </Emphasis>var. <Emphasis Type="Italic">angulisporus</Emphasis>

 Umbelopsis gibberispora is described as a new species in the genus Umbelopsis, Umbelopsidaceae, Mucorales. The species differs from others in this genus by ellipsoidal sporangiospores with unilaterally thickened walls. Phylogenetic analyses based on nuclear large subunit ribosomal DNA (nLSU rDNA) partial sequences suggest that U. gibberispora, U. swartii, and U. westeae form a clade together with the strains of Umbelopsis ramanniana. The ex-type strain of Micromucor ramannianus var. angulisporus is found to be very close to Umbelopsis vinacea, whereas other isolates identified under the former name in the sense of Linnemann fall in the U. ramanniana subclade. For these isolates, a new species, Umbelopsis angularis, is introduced. Phylogenetic relationships among Umbelopsis species are discussed related to their attributes of the sporangial wall and mature spore shapes. Received: August 27, 2002 / Accepted: March 11, 2003 Acknowledgments We thank Dr. Takashi Ohsono, Graduate School of Agriculture, Kyoto University, Japan, for providing the strain of U. gibberispora (CBS 109328). We also thank Dr. Wieland Meyer, University of Sydney, Australia for access to the phylogenetic tree based on ITS sequence data before publishing, and Dr. Richard C. Summerbell, Centraalbureau von Schimmelcultures, the Netherlands, for linguistic corrections.     

<Emphasis Type="Italic">Leifiporia rhizomorpha</Emphasis> gen. et sp. nov. and <Emphasis Type="Italic">L. eucalypti</Emphasis> comb. nov. in Polyporaceae (Basidiomycota)

A new poroid wood-inhabiting fungal genus, Leifiporia, is proposed, based on morphological and molecular evidence, which is typified by L. rhizomorpha sp. nov. The genus is characterized by an annual growth habit, resupinate basidiocarps with white to cream pore surface, a dimitic hyphal system with generative hyphae bearing clamp connections and branching mostly at right angles, skeletal hyphae present in the subiculum only and distinctly thinner than generative hyphae, IKI–, CB–, and ellipsoid, hyaline, thin-walled, smooth, IKI–, CB– basidiospores. Sequences of ITS and LSU nrRNA gene regions of the studied samples were generated, and phylogenetic analyses were performed with maximum likelihood, maximum parsimony and Bayesian inference methods. The phylogenetic analysis based on molecular data of ITS + nLSU sequences showed that Leifiporia belonged to the core polyporoid clade and was closely related to Diplomitoporus overholtsii and Lopharia cinerascens, and then grouped with Pycnoporus and Trametes. Further investigation was obtained for more representative taxa in the Polyporaceae based on nLSU sequences, in which the results demonstrated that the genus Leifiporia formed a monophyletic lineage with a strong support (100 % BS, 100 % BP, 1.00 BPP). Both morphological and molecular evidence confirmed the placement of the new genus in the core polyporoid clade. In addition, a new combination, Leifiporia eucalypti, is proposed based on examination of its type material and phylogeny.     

Molecular identification of <Emphasis Type="Italic">Aspergillus</Emphasis> and <Emphasis Type="Italic">Eurotium</Emphasis> species isolated from rice and their toxin-producing ability

Thirty milled rice samples were collected from retailers in 4 provinces of Malaysia. These samples were evaluated for Aspergillus spp, infection by direct plating on malt extract salt agar (MESA). All Aspergillus holomorphs were isolated and identified using nucleotide sequences of ITS 1 and ITS 2 of rDNA. Five anamorphs (Aspergillus flavus, A. oryzae, A. tamarii, A. fumigatus and A. nigef) and 5 teleomorphs (Eurotium rubrum, E. amstelodami, E. chevalieri, E. cristatum and E. tonophilum) were identified. The PCR-sequencing based technique for sequences of ITS 1 and ITS 2 is a fast technique for identification of Aspergillus and Eurotium species, although it does not work flawlessly for differentiation of Eurotium species. All Aspergillus and Eurotium isolates were screened for their ability to produce aflatoxin and ochratoxin A (OTA) by HPLC and TLC techniques. Only A. flavus isolate UPM 89 was able to produce aflatoxins B1 and B2.     

Molecular phylogeny and morphology of <Emphasis Type="Italic">Ophiocordyceps unituberculata</Emphasis> sp. nov. (Ophiocordycipitaceae), a pathogen of caterpillars (Noctuidae,Lepidoptera) from Yunnan,China

Ophiocordyceps unituberculata, a new insect pathogenic fungus from southwestern China, is described using molecular phylogenetic and morphological data. This fungus differs from other Ophiocordyceps species by its enormously long monophialidic conidiogenous cells and a large periclinal protuberance often growing near the apex of conidiogenous cells, single conidia (lanceolate to fusiform) embedded in mucous sheath, and much larger conidial dimensions. Four-locus (nrSSU, nrLSU, tef-1α and rpb1) and ITS data phylogenetic analyses show that O. unituberculata belongs to the Hirsutella nodulosa clade within the genus Ophiocordyceps of Ophiocordycipitaceae and is a separate clade from other allied species. Molecular phylogeny and morphology both strongly support the distinctiveness of this taxon. The interspecific relationships in the H. nodulosa clade are discussed.